Why Is FocalTuning a Superior Immunoassay Platform?
The FocalTuning immunoassay platform is a flow-based, bead-based multiplex system that enables high-precision intra-sample background correction at the individual analyte level.
Traditional immunoassays like ELISA rely on buffer-only blank wells as negative controls. However, these fail to account for matrix effects—nonspecific interactions from complex biological samples—that often lead to false positives. Platforms such as ImmunoCAP use non-target–coated sponges for background correction, but differences between the surface chemistry of target and non-target coatings often yield inconsistent and inaccurate background estimates.
In flow cytometry, isotype controls are commonly used to assess nonspecific antibody binding, yet they rarely mimic the true background behavior of target-specific capture antibodies. Similarly, Luminex and Meso Scale Discovery (MSD) platforms implement intra-well background subtraction, typically using a single control bead to correct for nonspecific signal across all analytes in the same well. This “one-control-for-all” approach fails to address analyte-specific background variability, potentially compromising quantitative accuracy.
The NOVEOS platform represents an improvement by incorporating dual bead types per well—target-coated and non-coated control beads—to correct for intra-sample background. However, NOVEOS data are collected on a per-well basis, limiting the statistical resolution and granularity of signal analysis.
In contrast, the FocalTuning platform pairs each target capture bead with a matched control bead within the same sample. This design eliminates the need for buffer-only blanks, isotype antibodies, or external target-negative samples. Instead, it enables analyte-specific background subtraction using intra-sample, per-analyte controls. Because the platform analyzes data on a per-bead basis, it offers substantially greater statistical power and reproducibility than traditional per-well systems.
In terms of sensitivity, the platform’s fluorescence-based limit of detection (LOD) reaches the low picogram-per-milliliter (pg/mL) range. Furthermore, if metal isotope tags (as used in mass cytometry) are employed in place of fluorophores, the LOD can be further improved, potentially enabling sub-picogram or even femtogram-level detection.
The FocalTuning platform supports both manual operation and automated, high-throughput processing, making it a versatile and scalable solution for both research and clinical diagnostics.